رایناک قادری

Different species of Salmonella are important pathogenic bacteria in animals and humans that often infect poultry flocks. Most Salmonella infections in humans are caused by eating foods contaminated with this bacterium. The aim of this study was to determine the Salmonella infection status of several poultry farms in Khuzestan province and to determine the serotypes and antimicrobial resistance pattern of isolated Salmonella. In the year 1392 total of 1829 samples including stool swabs, litter samples, feathers and eggshells were collected from a broiler mother farm, two laying mother hen farms and an incubator and cultured in enriching and selective media and detected using biochemical tests according to standard methods. In this study, 10 Salmonella isolates were isolated including serotypes enteritidis (3 cases), corvallis (3 cases), typhimurium (3 cases) and ruzizi (1 case) using Salmonella serum type tests. After performing antibiogram test with standard disk diffusion method, it was determined that 100% of the isolates were resistant to Colistin, Cefalexin and Cefazolin. Isolates were highly resistant to Ceftazidime, Nalidixic acid, Ampicillin, Ciprofloxacin, Nitrofurantoin, Amoxicillin, Neomycin, Streptomycin, Kanamycin, Ceftizoxime, Gentamicin and Tetracycline. All Salmonella isolates in this study were sensitive to Amikacin, Enrofloxacin, Ceftriaxone, Imipenem, Cefixime, Cefotaxime, Ofloxacin, Cefepime, Piperacillin, Florfenicol, Furazolidone, Chloramphenicol and Trimethoprim- Sulfamethoxazole.

Hemorrhagic septicemia (HS) is principally an infectious disease of respiratory tract caused by Pasteurella multocida in ruminants and birds resulting in huge economic repercussions. In the mid 1930s and early 1980s the Iranian buffalo (Pm Razi0001) and chicken (Pm Razi0002) strains of Pasteurella multocida were collected respectively from diseased animals. These strains have been used ever since for preparation of pasteurellosis vaccine at Razi institute. Multi Locus Sequence Typing (MLST) analysis is a standard method of genotyping for P. multocida. This was conducted to assess genomic structure of the vaccine P. multocida strains at Razi institute. Genomic material was prepared from freshly cultured strains through a simplified boiling protocol. KMT1-PCR was used to authenticate identity of strains as P. multocida. The Australian RIRDC-MLST typing system targeting 7 housekeeping genes of adk, est, pmi, zwf, mdh, gdh and pgi loci with few modifications was performed on strains. The Sequence Type (ST) and the corresponding clonal complex of the strains was determined by consulting the RIRDC-MLST database. Two STs were identified with ST352 (a subgroup of clonal complex CC122) and ST129 (a subgroup of clonal complex CC129) assigned for the buffalo (Pm Razi0001) and fowl (Pm Razi0002) strains, respectively. If using these strains as vaccine for all the past consecutive decades has had major impacts in population and epidemiology of P. multocida in this country, much more work is required.

In 1930's first whole cell pertussis vaccines became available to the public heralding a dramatic success in overcoming the global burden of the disease. To date only a handful of B. pertussis strains have been used by international/local pertussis vaccine manufacturers. Inevitable well-documented genetic changes in the world population of this pathogen have prompted serious questions on suitability of traditional vaccine strains protect human against currently circulating wild isolates of Bordetella pertussis.
Analyzing the genetic diversity within the most frequently-used vaccine strains of B. pertussis in the world
A recently developed multi locus variable number of tandem repeats analysis (MLVA) genotyping system along with a bioinforamtic piece of analysis was conducted on 11 strain/sub-strains of B137, B203 (10536), C393, Cs, E476, Tohama I, J445 (134), B202 and J446 (509) plus 2 sub-strains of 134 and 509 that are used at Razi institute for preparation of pertussis vaccine. In this study have used 6 individual loci of VNTR1, VNTR3a, VNTR3b, VNTR4, VNTR5 and VNTR6.
Findings: Six distinct genotypes were recognized among the examined strains by comparing our data with the Dutch MLVA databank. These were all new and not reported before in the database.
This observation reiterates on necessity for detection of predominant native strains to include in vaccine preparations suitable for different countries.

For almost half a century Mycobacterium avium subspecies paratuberculosis (MAP) 316F and MAP III & V strains have been used in preparation of paratuberculin at Razi institute. In order to characterize their genomic properties, a multi-approach strategy was employed to authenticate their identity (PCR-IS900 or PCR-F57), their type (PCR-Collins, PCR-gyrA or PCR-gyrB) and their genotype (MLVA-Thibault or SSR-Amonsin). Consequently, both strains proved to be MAP and from a cattle type. In MLVA the genotype was found to be INMV2, a type identical to that of the French MAP 316F Merial sub-strain. In explanation of these observations, the identical patterns of both studies strains in MLVA as well as SSR typing might be a case of coincidence or a an indication of a mutual ancestral clone. Besides, the fact that the supplier of MAP 316F strain studied here is the Turkish Etlik institute, it is likely that the original source for this strain is French. Application of the strategy developed in this study on indigenous MAP isolates will improve the current molecular epidemiology understanding of paratuberculosis in Iran.

Iran remains a major stronghold for glanders in the Middle East. In Iran, the non-indigenous Burkholderia mallei Razi 325 strain is used in manufacturing of the mallein, required for malleination of animals. Multi Locus Variable number tandem repeat analysis is currently the standard globally accepted genotyping system for Burkholderia mallei. This study was done to survey the genomic structure of Burkholderia mallei Razi 325, the strain used for industrial production of Mallein.
In this descriptive study, a MLVA genotyping system with 4 previously-characterized loci VNTR140, VNTR1367, VNTR2065, VNTR2971 along with two new loci of VNTR24, VNTR41 was used.
Optimization of PCRs resulted in a single protocol that enabled simultaneous amplification of all the six loci. Sequencing of PCR products revealed there were 2, 3, 12, 6, 1 and 2 copies of the unit repeat hold in the genome of the Burkholderia mallei Razi 325 strain. This observation was extended to include the already-whole genome sequenced Chinese Burkholderia mallei ATCC 23344 and Burkholderia mallei BMQ and also Burkholderia mallei SAVP1 strains.
The Burkholderia mallei Razi 325 strain is distinguishable from the other three strains through MLVA genotyping method.

L’objectif de cette étude était d’isoler, d’identifier et d’évaluer la susceptibilité antimicrobienne des agents responsables de la pasteurellose pneumonique chez les moutons de l’Est Azerbaïdjan, situé au nord ouest de l’Iran. Trois cent vingt (320) cas de pneumonie ont été détectés et le prélèvement des poumons affectés a été mené dans un abattoir. Tous les échantillons ont été ensuite soumis à des analyses bactériologiques et l’incidence de deux microorganismes spécifiques appartenant à la famille Pasteurellaceae a été étudiée. Selon nos analyses, six prélèvements pulmonaires étaient contaminés par Pasteurella multocida (1,87%), alors qu’aucun échantillon positif à la souche Mannheimia haemolytica n’a été détecté. Après l’isolation et la détection des microorganismes par des testes de culture et d’observation morphologiques, les souches bactériennes ont été identifiées grâce à leurs propriétés biochimiques et la méthode de réaction en chaîne par polymérase (PCR). La susceptibilité antimicrobienne de tous les isolats de P. multocida a été évaluée par la méthode de microdilution du bouillon de culture. L’évaluation de la concentration inhibitrice minimum (MIC) de 8 agents antimicrobiens vis-à-vis des isolats identifiés a révélé une résistance à l’ amoxicilline et une susceptibilité relative au ceftiofur. En conclusion, P. multocida se présente comme la cause majeure de pasteurellose pneumonique ovine dans la province de l’Est Azerbaïdjan et la fréquence cette épidémie varie de façon significative d’une saison à l’autre (P

Salmonella infections are the second leading cause of zoonotic bacterial foodborne illness. Main source of infection in human is contaminated food products. The aim of this study was sub typing isolates of Salmonella entericaobtained during our previous study byPulsed Field Gel Electrophoresis (PFGE) technique. All 46 Salmonella isolates were serotyped and then subjected to PFGE. Total isolates were analyzed by means of the molecular technique XbaI PFGE. In this study, PFGE and serotyping were used to subtype 46 Salmonella isolates belonging to 27different serovars and derived from human and different food origins. Among these isolates, S. Typhimurium was found to be the most predominant serovar. 40 PFGE patterns out of 46 isolates were obtained. The Discrimination Index obtained by serotyping (DI = 0.93) was lower than PFGE (DI = 0.99). Subtyping of Salmonella enterica is very important and shows that animal origin can be one of a reservoir that potentially could be transferred to human through the food chain. In addition, results of this study also revealed that this procedure is a golden standard for genotyping of such salmonellaserotypes.

Paratuberculosis caused by Mycobacterium avium subsp. paratuberculosis (Map) is a progressive, chronic and non-curable granulomatous enteritis. Paratuberculosis affects ruminants worldwide. The earliest reports on paratuberculosis in Iran date back to 1960’s when the disease was diagnosed in imported animals of the Abadan Oil Company’s farm. In the work presented here 13 field MAP isolates including 4 ovine and 3 caprine isolates from Isfahan, 6 bovine isolates from Zanjan and Alborz provinces plus 2 vaccinal strains of 316F and III & V were subjected to PCR-IS900, PCR-F57 for identification and also PCR-Collins for typing. In consequence, while an identical PCR protocol was successfully developed to simultaneously perform all the three experiments, the 15 tested bacteria were all shown to be MAP Type II (cattle type). In epidemiological sense, when the size of national goat, sheep and cattle herds of Iran is taken into account, circulation of MAP Type II is intriguing.

Salmonella enterica is recognized as one of the major food-borne pathogens with more than 2,500 serotypes worldwide. The present study addresses antimicrobial resistance of Salmonella enterica serovar Enteritidis isolates in Iran. A collection of 151 Salmonella spp. isolates collected from poultry were serotyped to identify Salmonella Enteritidis. Sixty-one Salmonella Enteritidis were subsequently tested against 30 antimicrobials. A high frequency of antimicrobial resistance was observed against nitrofurantoin (n=55, 90.2%) followed by nalidixic acid (n=41, 67.2%), and cephalexin (n=23, 37.7%). Multi drug resistance were observed in 35 (57.4%) out of 61 isolates. Twenty-six antimicrobial resistancepatterns were observed among the 61 Salmonella Enteritidis. All isolates were susceptible to ofloxacin, imipenem, enrofloxacin, chloramphenicol, gentamicin, and 3rd and 4th generation cephalosporins. In conclusion, our results revealed that implementing new policies toward overuse of antimicrobial drugs in Iranian poultry industry are of great importance.

The genus of Salmonella is very polymorphic and comprised of a number of genetically closely related serotypes. It is one of the emerging pathogen in food-borne disease which is often found in contaminated chicken eggs. Salmonella enterica is considered one of the major pathogens in public health worldwide. A total of 31 Salmonella isolates identified by specific antisera، which included Salmonella enteritidis (51. 6%)، Salmonella typhimurium (25. 8%)، Salmonella infantis (19. 4%) and Salmonella colindale (3. 2%). DNA was extracted using phenol- chloroform- isoamylalchol method. All the isolates showed fliC gene (1500bp) by using specific primers. PCR products were subjected to digestion using HhaI restriction endonuclease. PCR- RFLP results showed 3 patterns between all isolates. Our research gained in this study demonstrated that using HhaI restriction endonuclease could differentiate Salmonella enteritidis and Salmonella colindale but there is similarity between pattern of Salmonella typhimurium and Salmonella infantis.

SNP typing is now a well-established genotyping system in Bacillus anthracis studies. In the original standard method of Van Erth, SNPs at 13 loci of the B. anthracis genome were analyzed. In order to simplify and make appropriate this expensive method to low-budget laboratory settings, 13 primer pairs targeting the 13 corresponding SNPs were designed. Besides, a universal PCR protocol was developed to enable simultaneous amplification of all loci by conventional PCR machines. The efficiency of this approach was approved by applying on nine isolates of B. anthracis. We recommend using this modified procedure as an efficient alternative to Van Erth method until developing newer and affordable techniques.

Glanders caused by Burkholderia mallei, is a zoonotic disease. mallein test is the most used method in diagnosis of glanders. mallein is still prepared of standard B. mallei strain (s). In order to characterize the genomic properties, we have studied nucleotide structure of 23SrDNA, flip and BimA genes of a B.mallei strain that is used for preparation of mallein at Razi Institute and also a B.mallei isolate collected from a diseased tiger linked to the latest glanders outbreak at Tehran zoo. The isolated bacteria were cultivated on specific media. Bacterial suspensions were used to inoculate two guinea pigs in order to develop Strauss phenomenon. Genomic DNA was extracted by Isoamyl alcohol-Chloroform method. The PCR developed by using specific primers of B.mallei (three pairs) and common between B.mallei and B.pseoudomallei (one pair). The nucleotide sequence of the amplified genes was compared with standard fully sequenced genome available in gene bank. Both inoculated guinea pigs developed the characteristic Strauss phenomenon. Bacterial growth was the quantitative and qualitative different in the media and animal boudy. The amplificating four loci were successful. The nucleotide sequence analysis indicated these four loci were identical in B.mallei sterain/isolate invegtigented. This strategy represented a diagnostic algorithm suitable for differentiation between B.mallei and B.pseudomallei. Because importance of disease and limitation of mullein test in clinical and advanced cases of glanders using of sensitive and rapid diagnostic methods dependent on PCR and aknowledge of epidemiology molecular of glanders, is useful.

Within the course of this 18-month long work spanning 2010 and 2011, a total of eight human Mycobacterium bovis isolates collected from 1,320 tuberculosis-suspected patients, along with 68 bovine isolates were subjected to PCR-RFLP of pncA gene to assess their resistance to Pyrazynamide. All the isolates were subsequently spoligotyped to better understand their potential epidemiological links. As results showed, only 0.6% of the patients were infected with M. bovis, i.e. a considerably smaller rate compared to previous reports from Iran. Besides, all the isolates proved to be resistant to pyrazynamide. Consulting the SPOLDB4 spoligotyping databank, two patterns, namely ST595 and ST694, were detected among human isolates. Moreover, 12 patterns were found among the bovine isolates. ST595 was the single spoligotype shared between human and bovine hosts. For health officials, these observations indicate that in the Iranian environment M. bovis continues to produce a health risk to human.

Salmonella serotypes are important zoonotic pathogens in humans and animals. Most Salmonella infection in humans result from the ingestion of contaminated food. Resistance of Salmonella to commonly used antimicrobials is increasing and has emerged as a global problem. The main objective of this study was determine the frequency of serotypes of salmonella and antibiotic resistance of isolated Salmonella from poultry in Arak. A total of 245 samples were collected from poultry in Arak، and 75 Salmonella (30. 61%) were isolated. The result of Serotyping was Enteritidis (45. 33%)، Infantis (44%)، Typhimurium (5. 33%)، Bardo (2. 67%) and Bacongo (2. 67%). The prevalent Salmonella strains isolated in this study belonged to serotype Enteritidis and Infantis. The results of antibiogram test that was performed by the Kirby-Bauer disk diffusion method according to Clinical Laboratory Standards Institute (CLSI) showed that all isolated (100%) were sensitive to enrofloxacin، gentamicin، imipenem and ceftriaxone and the highest resistance were observed against nalidixic acid and nitrofuratoin. In addition، 63 isolates (84%) out of 75 had multi drug resistance to three or more antibiotics.